Department of Biotechnology, Faculty of Agricultural Sciences and Engineering, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.
Department of Genetics, Faculty of Biology, University of Seville, Seville 41012, Spain.
Can J Microbiol. 2021 May;67(5):406-414. doi: 10.1139/cjm-2020-0438. Epub 2020 Nov 23.
Fungal protoplast fusion is an approach to introduce novel characteristics into industrially important strains. Cellulases, essential enzymes with a wide range of biotechnological applications, are produced by many species of the filamentous fungi In this study, a collection of 60 natural isolates were screened for Avicel and carboxymethyl cellulose degradation, and two cellulase producers of and were used for protoplast fusion. One of the resulting hybrids with improved cellulase activity, C1-3, was fused with the hyperproducer Rut-C30. A new selected hybrid, F7, was increased in cellulase activity 1.8 and 5 times in comparison with Rut-C30 and C1-3, respectively. The increases in enzyme activity correlated with an upregulation of the cellulolytic genes , , , and in the parents. The amount of mRNA of and in F7 resembled that of Rut-C30 while the mRNA level was similar to that of C1-3. AFLP (amplified fragment length polymorphism) fingerprinting and GC-MS (gas chromatography - mass spectrometry) analysis represented variations in parental strains and fusants. In conclusion, the results demonstrate that a 3-interspecific hybrid strain was isolated, with improved characteristics for cellulase degradation and showing genetic polymorphisms and differences in the volatile profile, suggesting reorganizations at the genetic level.
真菌原生质体融合是一种向工业上重要菌株引入新特性的方法。纤维素酶是一种具有广泛生物技术应用的重要酶，许多丝状真菌都能产生。在这项研究中，对 60 个天然分离株进行了筛选，以检测它们对微晶纤维素和羧甲基纤维素的降解能力，然后使用两种纤维素酶产生菌 和 进行原生质体融合。其中一个具有改良纤维素酶活性的杂种 C1-3 与高产菌 Rut-C30 融合。一个新的选择杂种 F7 的纤维素酶活性分别比 Rut-C30 和 C1-3 提高了 1.8 倍和 5 倍。酶活性的增加与纤维素酶基因 的上调相关，这些基因在亲本中分别上调了 、 、 和 。F7 中的 和 的 mRNA 量与 Rut-C30 相似，而 的 mRNA 水平与 C1-3 相似。AFLP（扩增片段长度多态性）指纹图谱和 GC-MS（气相色谱-质谱）分析显示了亲本菌株和融合子的变化。总之，结果表明分离出了一种具有改良纤维素降解特性的 3 种属间杂交菌株，并表现出遗传多态性和挥发性成分的差异，提示在遗传水平上发生了重组。
